Titanium dioxide nanoparticles perturb the blood-testis barrier via disruption of actin-based cell adhesive function

Abstract

As one of the most commonly used nanoparticles, titanium dioxide nanoparticles (TiO₂-NPs) are widely used as coating reagents in cosmetics, medicine and other industries. The increasing risk of exposure to TiO₂-NPs raises concerns about their safety. In this study, we investigated the mechanism by which TiO₂-NPs cross the blood-testis barrier (BTB). TM-4 cells were selected as an <i>in vitro</i> Sertoli cell model of BTB. Cell viability, cell morphological changes, apoptosis, oxidative damage, and the expression levels of actin regulatory and tight junction (TJ) proteins were assessed in TM-4 cells treated with 3-nm and 24-nm TiO₂-NPs. Cells treated with 3-nm TiO₂-NPs exhibited increased cytotoxicity and decreased Annexin II expression, whereas cells treated with 24-nm TiO₂-NPs exhibited increased Arp 3 and c-Src expression. Both TiO₂-NPs induced significant oxidative stress, decreased the expression of TJ proteins (occludin, ZO-1 and claudin 5), damaged the TJ structure, and exhibited enlarged gaps between TM-4 cells. Our results indicated that both TiO₂-NPs crossed the BTB by disrupting actin-based adhesive junctions of TM-4 cells; however, apoptosis was not observed. Our results provide new insights into how TiO₂-NPs cross the BTB.